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Monobody-mediated alteration of enzyme specificity

Nature Chemical Biology volume 11, pages 762764 (2015) | Download Citation

Abstract

Current methods for engineering enzymes modify enzymes themselves and require a detailed mechanistic understanding or a high-throughput assay. Here, we describe a new approach where catalytic properties are modulated with synthetic binding proteins, termed monobodies, directed to an unmodified enzyme. Using the example of a β-galactosidase from Bacillus circulans, we efficiently identified monobodies that restricted its substrates for its transgalactosylation reaction and selectively enhanced the production of small oligosaccharide prebiotics.

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Acknowledgements

We acknowledge the use of the University of Chicago Genomics Core, which is supported by US National Institutes of Health grant P30CA014599 to the University of Chicago Comprehensive Cancer Center.

Author information

Affiliations

  1. Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois, USA.

    • Shun-ichi Tanaka
    • , Akiko Koide
    •  & Shohei Koide
  2. Frontier Research Department, Gifu R&D Center, Amano Enzyme, Inc., Gifu, Japan.

    • Shun-ichi Tanaka
    • , Tetsuya Takahashi
    • , Satoru Ishihara
    •  & Satoshi Koikeda

Authors

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Contributions

S.T. and S. Koide designed the research. S.T. and A.K. generated and characterized the monobodies. S.T. and T.T. analyzed effects of the monobodies on enzyme reactions. T.T. and S.I. conducted mutagenesis analysis of BgaD. S. Koikeda and S. Koide supervised the research. S.T. and S. Koide wrote the manuscript and the other authors commented on it.

Competing interests

S. Tanaka, T. Takahashi, S. Ishihara and S. Koikeda are employees of Amano Enzyme that manufactures the Biolacta enzyme. S. Tanaka, T. Takahashi and S. Koikeda are shareholders of Amano Enzyme. The University of Chicago has filed a patent application on technologies described in this paper.

Corresponding author

Correspondence to Shohei Koide.

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    Supplementary Results, Supplementary Figures 1–8 and Supplementary Table 1.

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DOI

https://doi.org/10.1038/nchembio.1896

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